Summary
Этот документ описывает простую технику, чтобы побудить аллоантигена конкретных анергии в человеческом мононуклеарных клеток периферической крови. Техника может быть применена в клинике для порождают не alloreactive клетки донора. Настой этих клеток может улучшить восстановления иммунитета и снижения токсичности после аллогенной трансплантации гемопоэтических стволовых клеток.
Protocol
Discussion
Disclosures
Нет конфликта интересов объявлены.
Acknowledgments
Materials
Name | Company | Catalog Number | Comments |
Ficoll-Hypaque PLUS | GE Healthcare | 17-1440-02 | |
RPMI 1640 | Invitrogen | 11875-093 | |
Hepes 1M | Invitrogen | 15630-080 | |
L-Glutamine 200mM | Invitrogen | 25030-081 | |
Gentamicin | Invitrogen | 15750-060 | |
Human AB serum (heat inactivated) | Gemini Bio Products | 100-512 | Use validated batches |
Complete Culture Media (500ml) | 440ml RPMI 1640, 50ml AB serum, 5ml Hepes, 5 ml Glutamine, 167uL Gentamicin | ||
Irradiator | GammaCell 1000 | ||
T-25 Cell Culture Flasks with gas permeable caps | Corning | 3056 | |
Humanized Monoclonal anti-B7.1 and anti B7.2 antibodies | See protocol text | ||
U bottomed 96 well culture plates | BD Biosciences | 353077 | |
Monoclonal CD3 antibody | Beckman Coulter Inc. | IM0178 | Reconstitute with 200ml distilled water |
Monoclonal CD28 antibody | Beckman Coulter Inc. | IM1376 | Reconstitute with 200ml distilled water |
CMV grade 2 antigen | Microbix | EL-01-02 | |
Tritiated Thymidine | NEN | NET027 | |
Scintillation Fluid | PerkinElmer, Inc. | 1205-440 | |
Harvester | Tomtec | ||
Printed Filtermat A | PerkinElmer, Inc. | 1450-421 | |
Filter Bag | PerkinElmer, Inc. | 1450-432 | |
1450 MicroBeta TriLux Scintillation Counter | Wallac | ||
null |
References
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